Journal Article

Universal Fluorescent Labeling (UFL) Method for Automated Microsatellite Analysis

Masashi Shimizu, Natsuki Kosaka, Takashi Shimada, Takemitsu Nagahata, Hironori Iwasaki, Hisaki Nagai, Tadayoshi Shiba and Mitsuru Emi

in DNA Research

Published on behalf of Kazusa DNA Research Institute

Volume 9, issue 5, pages 173-178
Published in print January 2002 | ISSN: 1340-2838
Published online January 2002 | e-ISSN: 1756-1663 | DOI: http://dx.doi.org/10.1093/dnares/9.5.173

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We have devised a novel method for automated microsatellite analysis using “universal” fluorescent labeling. This system is based on polymerase chain reactions driven by sequence-specific primers and a reporter primer labeled with a fluorescent dye at its 5′ end. The forward sequence-specific primer is designed with a tag region bearing no homology to any human genomic sequence. Complementary tag sequences act as templates for the 6-carboxyfluorescein-labeled reporter primer, and those products can be analyzed with an autosequencer. The results we achieved with this assay system were consistent with the results of conventional assays using radioisotope-labeled primers, and diagnosis required less time. Furthermore, the fluorescent-labeled reporter primer is “universal” in that it can be used with different sequence-specific primers designed to carry the appropriate tag sequence at their 5′-ends. Our observations suggest that the “universal” fluorescent labeling method is an efficient tool for analyzing sequence variations in human DNA.

Keywords: genomic alteration; fluorescent labeling; tagged primer; microsatellites

Journal Article.  0 words. 

Subjects: Genetics and Genomics

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